Difference between revisions of "Team:MIT/Experiments/miRNA/more experiments"

Line 18: Line 18:
 
<br>
 
<br>
 
<h2 style="color:#000000;text-decoration:underline"> Flow Cytometry Analysis </h2>
 
<h2 style="color:#000000;text-decoration:underline"> Flow Cytometry Analysis </h2>
−
<img src = "https://static.igem.org/mediawiki/2016/6/6d/T--MIT--sensor.PNG" style = 'padding: 5px'; width: 400px; height = 100px; float: center; border:5px;'>
+
<img src = "https://static.igem.org/mediawiki/2016/9/97/T--MIT--transient_transfection.PNG" style = 'padding: 5px'; width: 400px; height = 100px; float: center; border:5px;'>
 
<p></p>
 
<p></p>
 
<br>
 
<br>

Revision as of 20:39, 16 October 2016

Description of Experiments

General Experimental Set-Up and Data Analysis

miRNA Sensor

Read about here



Transient Transfection



Flow Cytometry Analysis



Sensitivity of miRNA Target Sites



Purpose

We want to understand the sensitivity of our miRNA target site constructs.


Set Up

We chose one miRNA target site, 451a, and designed and ordered siRNA complementary to the target site. We co-transfected tert-immortalized human endometrial stromal cells with our miRNA sensors and varying concentrations of siRNA then measured the red fluorescence output to gauge repression.


Results



When increasing siRNA-451a 0 to 1 nM, there is a 10 fold repression of red fluorescence. Saturation occurs at approximately 10 nM. More experiments would need to be done to understand the responsiveness between 0 to 1 nM.



miRNA Profile of tHESC

Purpose


Set Up


Results